Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , |
Tipo de documento: | Capítulo de livro |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/978-1-4939-7312-5_19 http://hdl.handle.net/11449/170161 |
Resumo: | Drug delivery is of paramount importance, since the drug needs to be delivered to a specific site, in adequate concentration, avoiding degradation in order to provide therapeutic efficacy. Different nanocarriers have been used over the years for this purpose and liposomes are well-established systems due to the high biocompatibility and the possibility to vehiculate both hydrophilic and lipophilic drugs. In order to circumvent the rapid clearance by the reticuloendothelial system and to avoid the healthy cells exposure to the drug, long circulating liposomes containing polyethyleneglycol (PEG) and functionalized liposomes for targeted delivery have been developed. Immunoliposomes consist of liposomes containing antibodies or antibody fragments attached at the membrane surface. This attachment can be performed using PEG lipids, containing a reactive terminal group such as maleimide and thiolated antibodies. Additionaly, the use of PEG chains as spacers increases antibody–antigen affinity, since the antibody is not shielded by the steric hindrance of PEG and also due to the correct orientation of antibodies for interaction with receptors on cell surface. In this chapter, we describe and discuss in details the protocol to prepare anti-epidermal growth factor receptor (anti-EGFR) and anti-human epidermal growth factor receptor 2 (anti-HER2) liposomes using cetuximab and trastuzumab as antibodies. We present the direct coupling method based on the maleimide thioether reaction for these immunoliposomes preparation and present some characterization steps and in vitro studies in cell culture which can be used for better understanding these nanocarriers. |
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Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bondCetuximabCovalent couplingDirect conjugationImmunoliposomesTargetingTrastuzumabDrug delivery is of paramount importance, since the drug needs to be delivered to a specific site, in adequate concentration, avoiding degradation in order to provide therapeutic efficacy. Different nanocarriers have been used over the years for this purpose and liposomes are well-established systems due to the high biocompatibility and the possibility to vehiculate both hydrophilic and lipophilic drugs. In order to circumvent the rapid clearance by the reticuloendothelial system and to avoid the healthy cells exposure to the drug, long circulating liposomes containing polyethyleneglycol (PEG) and functionalized liposomes for targeted delivery have been developed. Immunoliposomes consist of liposomes containing antibodies or antibody fragments attached at the membrane surface. This attachment can be performed using PEG lipids, containing a reactive terminal group such as maleimide and thiolated antibodies. Additionaly, the use of PEG chains as spacers increases antibody–antigen affinity, since the antibody is not shielded by the steric hindrance of PEG and also due to the correct orientation of antibodies for interaction with receptors on cell surface. In this chapter, we describe and discuss in details the protocol to prepare anti-epidermal growth factor receptor (anti-EGFR) and anti-human epidermal growth factor receptor 2 (anti-HER2) liposomes using cetuximab and trastuzumab as antibodies. We present the direct coupling method based on the maleimide thioether reaction for these immunoliposomes preparation and present some characterization steps and in vitro studies in cell culture which can be used for better understanding these nanocarriers.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)School of Pharmaceutical Sciences of Ribeirao Preto University of Sao Paulo, Av. do Cafe s/nCollege of Pharmacy The Ohio State University, Columbus, 500 W. 12th Ave.School of Pharmaceutical Sciences UNESP, Araraquara, Rodovia Araraquara-Jau, km. 1UNL CONICET FBCB Cell Culture LaboratorySchool of Pharmaceutical Sciences UNESP, Araraquara, Rodovia Araraquara-Jau, km. 1FAPESP: #2012/10388-3FAPESP: #2012/21513-3FAPESP: #2012/23764-3FAPESP: #2013/15134-2FAPESP: #2014/22451-7Universidade de São Paulo (USP)The Ohio State UniversityUniversidade Estadual Paulista (Unesp)Cell Culture LaboratoryPetrilli, RaquelEloy, Josimar O. [UNESP]Lee, Robert J.Lopez, Renata F. V.2018-12-11T16:49:33Z2018-12-11T16:49:33Z2018-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bookPart229-237http://dx.doi.org/10.1007/978-1-4939-7312-5_19Methods in Molecular Biology, v. 1674, p. 229-237.1064-3745http://hdl.handle.net/11449/17016110.1007/978-1-4939-7312-5_192-s2.0-85029726294Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMethods in Molecular Biology0,616info:eu-repo/semantics/openAccess2021-10-23T15:41:33Zoai:repositorio.unesp.br:11449/170161Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-05-23T11:11:12.979401Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
title |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
spellingShingle |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond Petrilli, Raquel Cetuximab Covalent coupling Direct conjugation Immunoliposomes Targeting Trastuzumab |
title_short |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
title_full |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
title_fullStr |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
title_full_unstemmed |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
title_sort |
Preparation of immunoliposomes by direct coupling of antibodies based on a thioether bond |
author |
Petrilli, Raquel |
author_facet |
Petrilli, Raquel Eloy, Josimar O. [UNESP] Lee, Robert J. Lopez, Renata F. V. |
author_role |
author |
author2 |
Eloy, Josimar O. [UNESP] Lee, Robert J. Lopez, Renata F. V. |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) The Ohio State University Universidade Estadual Paulista (Unesp) Cell Culture Laboratory |
dc.contributor.author.fl_str_mv |
Petrilli, Raquel Eloy, Josimar O. [UNESP] Lee, Robert J. Lopez, Renata F. V. |
dc.subject.por.fl_str_mv |
Cetuximab Covalent coupling Direct conjugation Immunoliposomes Targeting Trastuzumab |
topic |
Cetuximab Covalent coupling Direct conjugation Immunoliposomes Targeting Trastuzumab |
description |
Drug delivery is of paramount importance, since the drug needs to be delivered to a specific site, in adequate concentration, avoiding degradation in order to provide therapeutic efficacy. Different nanocarriers have been used over the years for this purpose and liposomes are well-established systems due to the high biocompatibility and the possibility to vehiculate both hydrophilic and lipophilic drugs. In order to circumvent the rapid clearance by the reticuloendothelial system and to avoid the healthy cells exposure to the drug, long circulating liposomes containing polyethyleneglycol (PEG) and functionalized liposomes for targeted delivery have been developed. Immunoliposomes consist of liposomes containing antibodies or antibody fragments attached at the membrane surface. This attachment can be performed using PEG lipids, containing a reactive terminal group such as maleimide and thiolated antibodies. Additionaly, the use of PEG chains as spacers increases antibody–antigen affinity, since the antibody is not shielded by the steric hindrance of PEG and also due to the correct orientation of antibodies for interaction with receptors on cell surface. In this chapter, we describe and discuss in details the protocol to prepare anti-epidermal growth factor receptor (anti-EGFR) and anti-human epidermal growth factor receptor 2 (anti-HER2) liposomes using cetuximab and trastuzumab as antibodies. We present the direct coupling method based on the maleimide thioether reaction for these immunoliposomes preparation and present some characterization steps and in vitro studies in cell culture which can be used for better understanding these nanocarriers. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-11T16:49:33Z 2018-12-11T16:49:33Z 2018-01-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/bookPart |
format |
bookPart |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/978-1-4939-7312-5_19 Methods in Molecular Biology, v. 1674, p. 229-237. 1064-3745 http://hdl.handle.net/11449/170161 10.1007/978-1-4939-7312-5_19 2-s2.0-85029726294 |
url |
http://dx.doi.org/10.1007/978-1-4939-7312-5_19 http://hdl.handle.net/11449/170161 |
identifier_str_mv |
Methods in Molecular Biology, v. 1674, p. 229-237. 1064-3745 10.1007/978-1-4939-7312-5_19 2-s2.0-85029726294 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Methods in Molecular Biology 0,616 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
229-237 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1803045787584692224 |